复合碳源强化反硝化及N<sub>2</sub>O减排的微生物组学机制

刘毅麟; 许允荣; 梁蕴瑶; 谢晓婧; 郑海欣; 袁婧; 陈丽萍; 韦朝海; 邱光磊

doi:10.13205/j.hjgc.202604009

复合碳源强化反硝化及N₂O减排的微生物组学机制

doi: 10.13205/j.hjgc.202604009

刘毅麟¹,
许允荣¹,
梁蕴瑶¹,
谢晓婧¹,
郑海欣¹,
袁婧¹,
陈丽萍¹,
韦朝海^1,2,3,
邱光磊^1,2,3,4, ,

1. 华南理工大学环境与能源学院, 广州 510006;
2. 工业聚集区污染控制与生态修复教育部重点实验室, 广州 510006;
3. 固体废物污染控制与资源化广东省重点实验室, 广州 510006;
4. 国家黄河流域生态保护和高质量发展联合研究中心, 北京 100012

基金项目:

国家自然科学基金项目（52270035，51808297）；广东省自然科学基金项目（2021A1515010494）；广东省珠江人才计划（2019QN01L125）；广州市重点研发计划（2023B03J1334）

详细信息

作者简介:
刘毅麟(2001—),男,硕士研究生,主要研究方向为水污染控制理论与技术。18770880970@163.com

通讯作者:
邱光磊(1984—),男,教授,主要研究方向为水污染控制理论与技术。qiugl@scut.edu.cn

计量
- 文章访问数: 1
- HTML全文浏览量: 0
- PDF下载量: 0
- 被引次数: 0
出版历程
- 收稿日期: 2026-03-17
- 网络出版日期: 2026-06-06
- 刊出日期: 2026-04-01

Microbiome mechanisms of composite carbon sources for enhancing denitrification and reducing N₂O emissions

1. School of Environment and Energy, South China University of Technology, Guangzhou 510006, China;
2. Key Laboratory of Pollution Control and Ecological Restoration in Industrial Clusters, Ministry of Education, Guangzhou 510006, China;
3. Guangdong Provincial Key Laboratory of Solid Wastes Pollution Control and Recycling, Guangzhou 510006, China;
4. National Joint Research Center for Ecological Conservation and High Quality Development of the Yellow River Basin, Beijing 100012, China

摘要

摘要: 污水处理厂生物脱氮过程普遍受到进水碳源不足的限制，外加碳源成为强化反硝化的重要手段。然而，传统单一碳源难以满足复杂微生物群落的代谢需求，导致脱氮效率和稳定性受到影响。复合碳源通过提供多种电子供体，可改善微生物代谢协同作用，但其微生物作用机制仍有待深入解析。以市政污水处理厂活性污泥为研究对象，通过批次反硝化实验，结合宏基因组和宏转录组技术，系统分析了反硝化微生物群落结构与不同碳源条件下功能基因转录特征。结果表明：与乙酸钠作为单一碳源时相比，复合碳源体系反硝化速率从（6.822±0.141） mg/（L·h）提高至（8.370±0.186） mg/（L·h），同时N₂O累积量显著降低约55%。宏基因组分析表明，Ottowia、Rubrivivax、Thauera和Zoogloea为主要反硝化菌。宏转录组分析结果显示，复合碳源显著上调反硝化关键功能基因转录，其中nirS、norB和nosZ的转录水平分别提高37.8%、27.4%和48.6%。此外，复合碳源促进了不同菌群形成互补的碳代谢策略，从而增进电子供体供应并提高反硝化效率。综合研究结果发现，复合碳源通过调控复杂微生物群落功能基因转录协同提升反硝化性能，这为污水处理厂碳源优化提供了重要的理论依据与指导。
- 反硝化 /
- 复合碳源 /
- N₂O排放 /
- 宏基因组 /
- 宏转录组
Abstract: Biological nitrogen removal in wastewater treatment plants （WWTPs） is commonly limited by insufficient influent carbon sources， thus making external carbon addition an important strategy for enhancing denitrification. However， conventional single carbon sources often fail to meet the metabolic demands of complex microbial communities， resulting in reduced nitrogen removal efficiency and stability. Composite carbon sources， by providing multiple electron donors， can improve metabolic cooperation among microorganisms， yet their underlying microbial mechanisms are still insufficiently understood. In this study， activated sludge from a municipal wastewater treatment plant was used to investigate the microbial mechanisms of composite carbon sources during denitrification. Batch denitrification experiments were conducted in combination with metagenomic and metatranscriptomic analyses to systematically characterize the microbial community structure and functional gene expression under composite carbon source conditions. The results showed that， compared with sodium acetate as the single carbon source， the composite carbon source system increased the denitrification rate from （6.822±0.141） mg/（L·h） to （8.370±0.186） mg/（L·h）， while reducing N₂O accumulation by approximately 55%. Metagenomic analysis revealed that Ottowia， Rubrivivax， Thauera， and Zoogloea were the dominant denitrifying genera. Metatranscriptomic results further demonstrated that the composite carbon sources significantly upregulated the transcription of key denitrification genes， with nirS， norB， and nosZ increasing by 37.8%， 27.4%， and 48.6%， respectively. In addition， the composite carbon sources promoted complementary carbon metabolic strategies among different microbial communities， thereby enhancing the supply of electron donors and improving denitrification efficiency. These findings indicate that composite carbon sources synergistically enhance denitrification performance by regulating microbial community structure and functional gene expression， providing a theoretical basis for carbon source optimization in WWTPs.
- denitrification /
- composite carbon sources /
- N₂O emissions /
- metagenomics /
- metatranscriptomics

HTML全文

参考文献(56)

[1]	Ministry of Ecology and Environment，the People’s Republic of China. Report on the State of the Ecology and Environment in China 2024［EB/OL］. 2025. https://english.mee.gov.cn/Resources/Reports/soe/index.shtml. 中华人民共和国生态环境部.《2024中国生态环境状况公报》［EB/OL］. 2025. https://www.mee.gov.cn/hjzl/sthjzk/zghjzkgb/.
[2]	HUANG C，LIU Q，LI Z L，et al. Relationship between functional bacteria in a denitrification desulfurization system under autotrophic，heterotrophic，and mixotrophic conditions［J］. Water Research，2021，188：116526.
[3]	WANG H L，LI J L，DONG Y H，et al. Research progress on the effect of carbon sources on biological denitrification of nitrogenous wastewater［J］. Journal of East China Normal University（Natural Science），2026（1）：110- 119. 汪翰林，李佳乐，董一慧，等. 碳源对含氮废水生物反硝化的影响研究进展［J］. 华东师范大学学报（自然科学版），2026（1）：110- 119.
[4]	WANG Y L，WEI Y X，LI X L，et al. Research progress on wastewater nitrogen removal technology［J］. Environmental Engineering，2010，28（S1）：119- 123. 王耀龙，魏云霞，李晓丽，等. 废水脱氮技术研究进展［J］. 环境工程，2010，28（增刊1）：119- 123.
[5]	JIANG J，LIANG D，HU Y. Solid slow-release carbon sources improve the simultaneous nitrification and denitrification processes in low carbon resource wastewater［J］. Bioresource Technology，2022，365：128148.
[6]	VINEYARD D，HICKS A，KARTHIKEYAN K G，et al. Economic analysis of electrodialysis，denitrification，and anammox for nitrogen removal in municipal wastewater treatment［J］. Journal of Cleaner Production，2020，262：121145.
[7]	LI J S. Study on the effects of different carbon sources and C/N ratio on system denitrification［D］. Wuhan：Wuhan University of Technology，2011. 李金诗. 不同碳源、C/N比对系统反硝化影响研究［D］. 武汉：武汉理工大学，2011.
[8]	AHMED S M，RIND S，RANI K. Systematic review：External carbon source for biological denitrification for wastewater［J］. Biotechnology and Bioengineering，2023，120（3）：642- 658.
[9]	CONSTANTIN H，FICK M. Influence of C-sources on the denitrification rate of a high-nitrate concentrated industrial wastewater［J］. Water Research，1997，31（3）：583- 589.
[10]	GU X，LENG J，ZHU J，et al. Influence mechanism of C/N ratio on heterotrophic nitrification- aerobic denitrification process［J］. Bioresource Technology，2022，343：126116.
[11]	WANG S Y，YIN F F，HOU H X，et al. Biological denitrification using methanol as an external carbon source［J］. Journal of Beijing University of Technology，2009，35（11）：1521- 1526. 王淑莹，殷芳芳，侯红勋，等. 以甲醇作为外碳源的生物反硝化［J］. 北京工业大学学报，2009，35（11）：1521- 1526.
[12]	HAGMAN M，NIELSEN J L，NIELSEN P H，et al. Mixed carbon sources for nitrate reduction in activated sludge-identification of bacteria and process activity studies［J］. Water Research，2008，42（6）：1539- 1546.
[13]	WANG Q，DENG Q S，WU W，et al. Operational diagnosis and carbon source optimization of yongchuan wastewater treatment plant based on mechanistic model［J］. Environmental Engineering，2022，40（6）：219- 225. 王骞，邓巧斯，吴畏，等. 基于机理模型的永川污水处理厂运行诊断与碳源优化［J］. 环境工程，2022，40（6）：219- 225.
[14]	CHEN J，TANG X，WU X，et al. Relating the carbon sources to denitrifying community in full-scale wastewater treatment plants［J］. Chemosphere，2024，361：142329.
[15]	PAN Y，SUN R Z，WANG Y，et al. Carbon source shaped microbial ecology，metabolism and performance in denitrification systems［J］. Water Research，2023，243：120330.
[16]	ALAMUSI，CAI B J，WANG F，et al. Effects of carbon sources on the denitrification process and bacterial community in wastewater treatment［J］. Sichuan Environment，2015，34（1）：13- 18. 阿拉木斯，蔡碧婧，王峰，等. 碳源对污水处理反硝化过程及细菌种群的影响［J］. 四川环境，2015，34（1）：13- 18.
[17]	RAMÍREZ-MELGAREJO M，GASSÓ-DOMINGO S，GÜERECA L P. Evaluation of N₂O emissions in wastewater treatment systems:a comparative analysis of emission between case studies of developed and developing countries［J］. Water，Air，& Soil Pollution，2019，230（2）：42.
[18]	DAELMAN M R J，VAN VOORTHUIZEN E M，VAN DONGEN L G J M，et al. Methane and nitrous oxide emissions from municipal wastewater treatment– results from a long-term study［J］. Water Science and Technology，2013，67（10）：2350- 2355.
[19]	DUAN H，van den AKKER B，THWAITES B J，et al. Mitigating nitrous oxide emissions at a full-scale wastewater treatment plant［J］. Water Research，2020，185：116196.
[20]	ZHU G，SHI H，ZHONG L，et al. Nitrous oxide sources，mechanisms and mitigation［J］. Nature Reviews Earth& Environment，2025，6（9）：574- 592.
[21]	PAN Y，HUA T W，SUN R Z，et al. Machine learning-assisted optimization of mixed carbon source compositions for high-performance denitrification［J］. Environmental Science& Technology，2024，58（28）：12498- 12508.
[22]	WANG J H. Study on greenhouse gas emissions from municipal wastewater treatment plants［D］. Ji'nan：Shandong University，2012. 王金鹤. 城镇污水处理厂中温室气体的释放研究［D］. 济南：山东大学，2012.
[23]	HYATT D，CHEN G L，LOCASCIO P F，et al. Prodigal：prokaryotic gene recognition and translation initiation site identification［J］. BMC Bioinformatics，2010，11（1）：119.
[24]	LI W，GODZIK A. Cd-hit：a fast program for clustering and comparing large sets of protein or nucleotide sequences［J］. Bioinformatics，2006，22（13）：1658- 1659.
[25]	WANG Z J，WANG S，LIU Y Y，et al. Application of metagenomic technology in molecular detection of functional microorganisms involved in nitrogen cycle［J］. Biotechnology Bulletin，2018，34（1）：1- 14. 王朱珺，王尚，刘洋荧，等. 宏基因组技术在氮循环功能微生物分子检测研究中的应用［J］. 生物技术通报，2018，34（1）：1- 14.
[26]	LOU H C，XU X J，WANG L，et al. Study on Enhanced nitrogen removal efficiency of wastewater by hydrolysis denitrification + AO process［J］. Environmental Engineering，2018，36（12）：75- 80. 娄红春，许晓杰，王亮，等. 水解反硝化+AO工艺强化污水脱氮效能研究［J］. 环境工程，2018，36（12）：75- 80.
[27]	GRABHERR M G，HAAS B J，YASSOUR M，et al. Full-length transcriptome assembly from RNA-Seq data without a reference genome［J］. Nature Biotechnology，2011，29（7）：644- 652.
[28]	MA H，MA X G. Research progress on the application of metatranscriptomics in environmental microbiology［J］. Environmental Science and Technology，2026，39（1）：65- 69. 马欢，马兴冠. 宏转录组学在环境微生物应用中的研究进展［J］. 环境科技，2026，39（1）：65- 69.
[29]	Ministry of Environmental Protection. Water quality-determination of nitrate nitrogen-ultraviolet spectrophotometry：HJ/T 346—2007［S］. Beijing：China Environmental Science Press. 2007. 环境保护部. 水质硝酸盐氮的测定紫外分光光度法：HJ/T 346—2007［S］. 北京：中国环境科学出版社，2007.
[30]	Ministry of Environmental Protection. Water quality-determination of nitrite nitrogen-spectrophotometry：GB 7493—87［S］. Beijing：China Environmental Science Press. 1987. 环境保护部. 水质亚硝酸盐氮的测定分光光度法：GB 7493—87［S］. 北京：中国环境科学出版社 1987.
[31]	Ministry of Environmental protection. water quality-determination of ammonia nitrogen-nessler's reagent spectrophotometry：HJ 535—2009［S］. Beijing：China Environmental Science Press. 2009. 环境保护部. 水质氨氮的测定纳氏试剂分光光度法：HJ 535—2009［S］. 北京：中国环境科学出版社. 2009.
[32]	Ministry of Environmental Protection. Technical requirements for water quality automatic analyzer for total organic carbon（TOC）：HJ/T 104—2003［S］. Beijing：China Environmental Science Press. 2003. 环境保护部. 总有机碳（TOC）水质自动分析仪技术要求：HJ/T 104—2003［S］. 北京：中国环境科学出版社. 2003.
[33]	Ministry of Environmental Protection. Water quality-determination of suspended substance-gravimetric method：GB 11901—1989［S］. Beijing：China Environmental Science Press. 1989. 环境保护部. 水质悬浮物的测定重量法：GB 11901—1989［S］. 北京：中国环境科学出版社. 1989.
[34]	LOU C N，HAN X S，JIN Y，et al. Effects of different carbon sources on microbial denitrification performance［J］. Journal of East China University of Science and Technology（Natural Science Edition），2023，49（5）：660- 669. 楼超楠，韩昫身，金艳，等. 不同碳源对微生物反硝化性能的影响［J］. 华东理工大学学报（自然科学版），2023，49（5）：660- 669.
[35]	LI F，YU C Z，ZHANG W，et al. Research progress on enhancing biological denitrification using slow-release carbon sources［J］. Water Purification Technology，2023，42（8）：48- 58. 李芳，于成志，张唯，等. 利用缓释碳源强化生物反硝化脱氮的研究进展［J］. 净水技术，2023，42（8）：48- 58.
[36]	ZHENG Y H，REN Y Y，WANG Z B，et al. Performance of different acidogenic food waste fermentation liquids as carbon sources for wastewater denitrification［J］. Transactions of the Chinese Society for Agricultural Machinery，2024，55（11）：446- 452. 郑永辉，任玉莹，王珍宝，等. 不同产酸类型餐厨酸化液作为污水反硝化碳源的性能研究［J］. 农业机械学报，2024，55（11）：446- 452.
[37]	CHAI W Y，GUO Y N，YANG Z，et al. Research progress on the influence of solid carbon source characteristics on biological denitrification technology［J］. Journal of Environmental Engineering Technology，2024，14（3）：963- 972. 柴文云，郭亚南，杨铮，等. 固相碳源的特性对生物反硝化脱氮技术的影响研究进展［J］. 环境工程技术学报，2024，14（3）：963- 972.
[38]	ZUMFT W G. Cell biology and molecular basis of denitrification［J］. Microbiology and Molecular Biology Reviews，1997，61（4）：533- 616.
[39]	DELGADO-BAQUERIZO M，MAESTRE F T，REICH P B，et al. Microbial diversity drives multifunctionality in terrestrial ecosystems［J］. Nature Communications，2016，7（1）：10541.
[40]	GRAF D R H，JONES C M，HALLIN S. Intergenomic Comparisons Highlight Modularity of the Denitrification Pathway and Underpin the Importance of Community Structure for N₂O Emissions［J］. PLoS ONE，2014，9（12）：e114118.
[41]	KONOPKA A，LINDEMANN S，FREDRICKSON J. Dynamics in microbial communities:unraveling mechanisms to identify principles［J］. The ISME Journal，2015，9（7）：1488- 1495.
[42]	LOUCA S，POLZ M F，MAZEL F，et al. Function and functional redundancy in microbial systems［J］. Nature Ecology& Evolution，2018，2（6）：936- 943.
[43]	HEROLD M，MARTÍNEZ ARBAS S，NARAYANASAMY S，et al. Integration of time-series meta-omics data reveals how microbial ecosystems respond to disturbance［J］. Nature Communications，2020，11（1）：5281.
[44]	ZHANG T，SHAO M F，YE L. 454 Pyrosequencing reveals bacterial diversity of activated sludge from 14 sewage treatment plants［J］. The ISME Journal，2012，6（6）：1137- 1147.
[45]	OYSERMAN B O，NOGUERA D R，DEL RIO T G，et al. Metatranscriptomic insights on gene expression and regulatory controls in Candidatus Accumulibacter phosphatis［J］. The ISME Journal，2016，10（4）：810- 822.
[46]	RICHARDSON D J，BERKS B C，RUSSELL D A，et al. Functional，biochemical and genetic diversity of prokaryotic nitrate reductases［J］. Cellular and Molecular Life Sciences，2001，58（2）：165- 178.
[47]	PAN Y，NI B J，BOND P L，et al. Electron competition among nitrogen oxides reduction during methanol-utilizing denitrification in wastewater treatment［J］. Water Research，2013，47（10）：3273- 3281.
[48]	HALLIN S，PHILIPPOT L，LÖFFLER F E，et al. Genomics and Ecology of Novel N₂O-Reducing Microorganisms［J］. Trends in Microbiology，2018，26（1）：43- 55.
[49]	WANG S，LU Y P，LIU S Y，et al. Research on carbon emissions from suzhou wastewater treatment plants［J］. Environmental Engineering，2023，41（10）：173- 184. 王朔，陆云平，刘树洋，等. 苏州市污水处理厂碳排放研究［J］. 环境工程，2023，41（10）：173- 184.
[50]	CAO S，CHENG Z，KOCH K，et al. Municipal wastewater driven partial-denitrification（PD）aggravated nitrous oxide（N₂O）production［J］. Journal of Cleaner Production，2024，434：139916.
[51]	GÖRKE B，STÜLKE J. Carbon catabolite repression in bacteria：many ways to make the most out of nutrients［J］. Nature Reviews Microbiology，2008，6（8）：613- 624.
[52]	WANG D X，WU P K，LIANG L M，et al. Formation mechanisms and mitigation strategies of nitrous oxide in biological wastewater treatment：A review［J］. Chinese Journal of Applied and Environmental Biology，2023，29（2）：281- 288. 王东旭，伍佩珂，梁兰梅，等. 污水生物处理过程氧化亚氮的形成机理及减量策略研究进展［J］. 应用与环境生物学报，2023，29（2）：281- 288.
[53]	KOVÁROVÁ-KOVAR K，EGLI T. Growth Kinetics of Suspended Microbial Cells:From Single-Substrate-Controlled Growth to Mixed-Substrate Kinetics［J］. Microbiology and Molecular Biology Reviews，1998，62（3）：646- 666.
[54]	LU H，CHANDRAN K，STENSEL D. Microbial ecology of denitrification in biological wastewater treatment［J］. Water Research，2014，64：237- 254.
[55]	ZHANG Z，ZHAO M X，RUAN W Q. Study on the effects of different carbon sources on the short-cut nitrification and denitrification of food wastewater treatment［J］. Environmental Engineering，2018，36（7）：46- 50. 张周，赵明星，阮文权. 不同碳源对餐厨废水短程硝化反硝化处理效果的影响研究［J］. 环境工程，2018，36（7）：46- 50.
[56]	LU W B，HU H Y，FENG Y N，et al. Denitrification performance of polycaprolactone/corncob composite solid carbon source under different influent nitrate concentrations and temperatures［J］. Progress in Fishery Sciences，2024，45（2）：123- 135. 卢伟斌，胡海燕，冯宇娜，等. 不同进水硝酸盐浓度和温度下聚己内酯/玉米芯复合固体碳源的反硝化性能探究［J］. 渔业科学进展，2024，45（2）：123- 135.